Dimethoxy quinazoline compound and application thereof

Abstract

The invention belongs to the technical field of biochemistry, and particularly relates to a dimethoxy quinazoline compound and application thereof. Through innovative design and chemical structure adjustment, more effective non-small cell lung cancer resisting activity is provided; meanwhile, through innovative design and chemical structure adjustment, the problem of complex drug resistance generated by the EGFR tyrosine kinase inhibitor in the treatment process is solved in a targeted manner, and the anti-cancer cell proliferation activity of the drug in drug-resistant cells is improved; methoxyl groups are introduced into 6 and 7 sites in a quinazoline skeleton to increase the combination of the compound to a target spot, and a 4-methyl piperazine ethylamine side chain is introduced, so that potential sites of a chemical structure are fully utilized, the activity of the inhibitor to C797S-containing three-mutation EGFR is improved, and the blank in the prior art is filled.

Description

Technical Field

[0001] The invention belongs to the technical field of biochemistry, and specifically relates to a dimethoxyquinazoline compound substituted with a piperazine derivative and application thereof. Background Art

[0002] The incidence and mortality of lung cancer are among the highest in the world. Non-small cell lung cancer (NSCLC) is the most common type (accounting for 80% to 85%), and at least 70% of NSCLC patients are already in the advanced stage when they seek medical treatment, missing the best time for surgical treatment (Kuang Chen, Chen Min, Mao Mian, et al. Systematic review of icotinib in the treatment of advanced non-small cell lung cancer [J]. China Pharmacy, 2016, 27(9): 1219-1222.). Radiotherapy and chemotherapy are usually used clinically for such patients. Most traditional chemotherapy kills tumor cells by destroying DNA function and structure, interfering with transcription, translation and mismatch repair, and disrupting cell mitosis (Malhotra V, Perry MC. Classical chemotherapy: mechanisms, toxicities and the therapeutic window [J]. Cancer Biol Ther, 2003, 2: 2-4.). Since there is no specific target, chemotherapy drugs can kill or inhibit all rapidly proliferating cells, and will kill rapidly proliferating normal cells while killing tumor cells. Therefore, although it can prolong the survival time of advanced patients, it often produces many clinical adverse reactions due to cytotoxic effects, including bone marrow suppression, nausea, vomiting, diarrhea, mucositis, alopecia, ototoxicity, nephrotoxicity, etc. (ZRAIK IM, HEβ-BUSCHY. Management of chemotherapy side effects and their long-term sequelae [J]. Urologe A, 2021, 60 (7): 862-871.), and the 5-year survival rate is low (WADOWSKA K, BIL-LULA I, TREMBECKI, et al. Genetie markers in lung cancer diagnosis; areview [J]. Int J Mol Sci, 2020. 21 (13): 4569.). Targeted drugs are based on the biological differences between normal cells and tumor cells (including genes, protein kinases, and signal transduction pathways, etc.), inhibiting the growth and spread of tumors by affecting specific genes or proteins, with no effect or little effect on normal cells, and are ideal drugs for individualized treatment. Receptor tyrosine kinase activity is strictly regulated in normal cells, but in tumor cells, due to mutation or abnormal expression, its kinase activity and its downstream signal transduction pathways are often abnormally activated, promoting the occurrence and progression of tumors.

[0003] Epidermal growth factor receptor (EGFR) is a transmembrane transporter with tyrosine kinase activity. It regulates a variety of cell life activities including proliferation, differentiation, apoptosis and migration by triggering the activation of intracellular signaling pathways (Shi Y Au JS, Thongprasert S, et al. A prospective, molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology (PIONEER) [J]. Thorac Oncol, 2014, 9 (2): 154-162.). EGFR gene mutations change the spatial structure of its encoded protein, resulting in a higher affinity for the intracellular ATP binding domain of EGFR. Abnormal activation of the kinase structure will lead to uninhibited cell growth, ultimately promoting the occurrence and metastasis of tumors. In NSCLC patients, EGFR gene mutation is the most frequent targetable driver mutation (Cirri A, Yarden Y. EGF-ERBB signalling: towards the systems level [J]. Nat Rev Mol Cell Biol, 2006, 7 (7): 505-516.), and the mutation rate in Asian populations exceeds 50% (Hynes NE, Lane HA. ERBB receptors and cancer: the complexity of targeted inhibitors [J]. Nat Rev Cancer, 2005, 5 (5): 341-354.), especially in non-smokers and women.EGFR gene mutations mainly occur in exons 18-21 of the EGFR tyrosine kinase coding region, among which exon 19 deletion (ex19del) and exon 21 L858R point mutation are the most common EGFR mutations, also known as EGFR sensitive mutations (Murray S, Dahabreh IJ, Linardou H, et al. Somatic mutations of the tyrosine kinase domain of epidermal growth factor receptor and tyrosine kinase inhibitor response to TKIs in non-small-cell lung cancer: all analytical database [J]. Thorac Oncol, 2008, 3 (8): 832-839.). Exon 20 insertion (ex20ins) mutation is the most common non-classical EGFR gene mutation, which is more common in women, non-smokers and patients with lung adenocarcinoma. Among them, the incidence of EGFR20 exon insertion mutation accounts for about 4% to 12% of all EGFR mutation NSCIC patients, which is second only to EGFR19 exon deletion mutation and EGFR21 exon L858R mutation. In recent years, with the continuous development of molecular targeted research, EGFR-tyrosine kinase inhibitors (TKIs) have become the main therapeutic drugs for the treatment of EGFR gene mutation-positive NSCLC (OHMORI T, YAMAOKA T, ANDO K, et al. Molecular and clinical features of EGFR-TKI-associated lung injury [J]. Int J Mol Sci, 2021, 22 (2): 792). EGFR-TKIs compete with ATP for the active zone of tyrosine kinase, inhibit tyrosine kinase phosphorylation, and block downstream signal transduction pathways to achieve the effect of tumor inhibition.

[0004] Compared with traditional chemotherapy, EGFR-TKIs have precise efficacy, mild adverse reactions and are easy to use. The first generation of EGFR-TKI is a class of 4-anilinoquinazoline compounds with quinazoline as the parent core. Representative drugs include gefitinib, erlotinib, and icotinib. It is a reversible, non-selective tyrosine kinase inhibitor that has inhibitory effects on both wild-type and mutant EGFR (Exl9Del point mutation and 21L858R substitution mutation). However, patients develop acquired resistance to these inhibitors after 9 to 13 months. Common EGFR mutations associated with drug resistance include exon 19 deletion, L858R mutation and T790M mutation, and about 50% to 60% of drug resistance is due to EGFR T790M mutation (KOBAYASHI S, BOGGON TJ, DAYARAM T, et al EGFR mutation and resistance of non-small-cell lung cancer to gefitinib [J]. The New England Journal of Medicine, 2005, 352 (8): 786-792.). Unlike the first-generation EGFR-TKI, the second-generation EGFR-TKI is an aminocrotonylamino-substituted quinazoline derivative, which binds to the tyrosine kinase activation domain in a non-selective and irreversible manner. Representative drugs include afatinib and dacomitinib. However, since it is a pan-EGFR inhibitor, it can simultaneously inhibit the phosphorylation process of multiple receptors such as EGFR, HER-2 and HER-4, and has the characteristics of multiple targets and strong adverse reactions (Bao R, Gao E Zhang F, et al. Abslract 3063: Discovery of a third-generation EGFR inhibitor, which is highly selective and potent against both resistant and activating EGFR mutations for NSCLC [J]. Therapy Cancer Res, 2016, 76: 3063-3063.). Unlike the first- and second-generation EGFR-TKIs that use quinazoline as the parent ring, the third-generation EGFR-TKI uses a pyrimidine ring as the parent ring structure, while retaining the Michael addition receptor-acrylamide structure, forming a hydrogen bond with Met793 in the EGFR ATP binding pocket through the pyrimidine parent ring, and forming a covalent bond with the EGFR cysteine ​​residue Cys797 through acrylamide to inhibit the conduction of the signal pathway, and this binding is selective and irreversible. This enables it to not only treat EGFR gene sensitive mutations, but also solve the problem of T790M resistance mutations produced by previous generation EGFR-TKIs.Moreover, the third-generation EGFR-TKI is highly selective and has a weaker inhibitory effect on wild-type EGFR than the first- and second-generation EGFR-TKIs, so the drug has less toxic side effects (ShiYHu X, Zhang S, et al. Efficacy, safety, and genetic analysis of furmonertinib (AST2818) in patients with EGFR T790M mutated non-small-cell lung cancer: a phase 2b, multicentre, single-arm, open-label study [J]. The Lancet Respiratory Medicine, 2021, 9 (8): 829-839.). Representative drugs include osimertinib, aumelinib, furmonertinib, and befotertinib. The fourth-generation drug is still in the clinical trial stage.

[0005] Although the marketed EGFR inhibitors have been successful in treatment, they still have some limitations, including insufficient inhibition of triple-mutant EGFR and drug resistance in some patients. For patients with EGFR mutation-positive NSCLC, the use of third-generation EGFR-TKI treatment has prolonged the progression-free survival of the disease, and the objective response rate and disease control rate have been significantly improved. However, after a period of treatment with third-generation EGFR-TKI, almost all patients will develop third-generation EGFR-TKI resistance, which greatly affects the treatment effect of patients with advanced EGFR mutation-positive NSCLC (Soria JC, Ohe Y, Vansteenkiste J et al. Osimertinib in Untreated EGFR-Mutated Advanced Non-Small-Cell Lung Cancer[J]. N Engl J Med 2018; 378: 113-125; Papadimitrakopoulou VA, Mok TS, Han JY et al. Osimertinib versus platinum-pemetrexed for patients with EGFR T790M advanced NSCLC and progression on a prior EGFR-tyrosine kinase inhibitor: AURA3 overall survival analysis[J]. Ann Oncol 2020;31:1536-1544.). Therefore, in order to prolong the survival of patients and improve their quality of life, it is very necessary to develop new inhibitors against third-generation EGFR-TKI resistance. Summary of the invention

[0006] The present invention obtains a new 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound through innovative design and chemical structure adjustment. The compound has a wider spectrum of activity and is more effective, especially for complex EGFR variant non-small cell lung cancer. Based on this, the present invention is completed.

[0007] In the first aspect, the present invention provides a 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound or a pharmaceutically acceptable salt thereof; the compound structural formula is shown in (I):

[0008]

[0009] Wherein, R is selected from a halogen atom, an aryl group or a heteroaryl group.

[0010] Furthermore, the aromatic group may be a benzene ring or an aromatic hetero group.

[0011] Furthermore, the aryl group and the aromatic hetero group are substituted by one or more substituents.

[0012] Furthermore, the substituent includes, but is not limited to, an alkyl group, a phenolic hydroxyl group or a halogen.

[0013] In one embodiment of the present invention, the structural formula of the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound may be:

[0014]

[0015] Further, the pharmaceutically acceptable salts of the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound include but are not limited to hydrochloride, sulfate, maleate, succinate, adipate, glycolate, malate, fumarate, benzenesulfonate, benzoate, hippurate or oxalate.

[0016] Furthermore, the pharmaceutically acceptable salt of the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound also includes solvates, hydrates or polymorphs of the above salts.

[0017] In a second aspect, the present invention provides a method for preparing the compound of the first aspect, the method comprising the following steps:

[0018] S1. Using 2,4-dichloro-6,7-dimethoxyquinazoline as a raw material, 2-(4-methylpiperazine-1-yl)ethylamine was added to carry out a nucleophilic substitution reaction on the chlorine at the 4 position;

[0019] S2. Add substituted aryl boronic acid to the reaction system of S1, and use a metal catalyst to carry out a Suzuki coupling reaction to obtain the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound.

[0020] Furthermore, in step S1, the 2,4-dichloro-6,7-dimethoxyquinazoline and 2-(4-methylpiperazine-1-yl)ethylamine are added in a molar ratio of 1:1.2.

[0021] Furthermore, in step S1, the nucleophilic substitution reaction temperature is 15-40°C, preferably 25°C.

[0022] Furthermore, in step S1, the nucleophilic substitution reaction time is 24-48 hours, preferably 48 hours.

[0023] Further, in step S2, the substituted aromatic boronic acid includes but is not limited to benzo[d][1,3]dioxane-4-ylboronic acid, (5-fluoro-2-hydroxyphenyl)boronic acid, 3-furylboronic acid, (5-chloro-2-hydroxyphenyl)boronic acid, 2-hydroxyphenylboronic acid, 3-hydroxynaphthalen-2-ylboronic acid, phenylboronic acid, 3,5-dichlorophenylboronic acid, 2,5-dimethoxyphenylboronic acid or 5-cyano-2-methoxyphenylboronic acid).

[0024] Furthermore, in step S2, the substituted aryl boronic acid and the metal catalyst are added in a molar ratio of 1:0.01.

[0025] Further, in step S2, the metal catalyst includes but is not limited to palladium tetrakistriphenylphosphine, palladium acetate, dichlorobis(triphenylphosphine)palladium, di(tri-tert-butylphosphine)palladium or palladium on carbon (Pd / C).

[0026] Further, in step S2, the Suzuki coupling reaction temperature is 100-130°C, preferably 115°C.

[0027] Further, in step S2, the Suzuki coupling reaction time is 5-10 hours, preferably 8 hours.

[0028] Furthermore, in step S2, triethylamine, n-butanol and toluene need to be added to react together during the Suzuki coupling reaction.

[0029] In a third aspect, the present invention provides a pharmaceutical composition, comprising the compound represented by formula (I) of the first aspect or a pharmaceutically acceptable salt thereof and an optional pharmaceutically acceptable carrier; the compound structural formula is shown in (I):

[0030]

[0031] Wherein, R is selected from a halogen atom, an aryl group or a heteroaryl group.

[0032] Furthermore, the aromatic group may be a benzene ring or an aromatic hetero group.

[0033] Furthermore, the aryl group and the aromatic hetero group are substituted by one or more substituents.

[0034] Furthermore, the substituent includes, but is not limited to, an alkyl group, a phenolic hydroxyl group or a halogen.

[0035] Furthermore, pharmaceutically acceptable salts of the compound represented by formula (I) include, but are not limited to, hydrochloride, sulfate, maleate, succinate, adipate, glycolate, malate, fumarate, benzenesulfonate, benzoate, hippurate or oxalate.

[0036] Furthermore, the pharmaceutically acceptable salts of the compound represented by formula (I) also include solvates, hydrates or polymorphs of the above salts.

[0037] Furthermore, the carrier material includes, but is not limited to, one or more of a water-soluble carrier material, a poorly soluble carrier material and / or an enteric carrier material.

[0038] Furthermore, the water-soluble carrier material includes, but is not limited to, one or more of polyethylene glycol, polyvinyl pyrrolidone and / or organic acid.

[0039] Furthermore, the poorly soluble carrier material includes but is not limited to one or more of ethyl cellulose and / or cholesterol stearate.

[0040] Furthermore, the enteric carrier material includes but is not limited to one or more of cellulose acetate phthalate and / or carboxymethyl ethyl cellulose.

[0041] Furthermore, the pharmaceutical composition can be prepared into a variety of dosage forms, including but not limited to one or more of tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories and / or lyophilized powder injections.

[0042] Furthermore, the preparation can be one or more of a conventional preparation, a sustained-release preparation, a controlled-release preparation and / or various microparticle delivery systems.

[0043] Furthermore, the tablets may widely use various carriers known in the art, including one or more of diluents and absorbents, wetting agents and binders, disintegrants, disintegration inhibitors, absorption promoters and / or lubricants.

[0044] Furthermore, the diluent and absorbent include, but are not limited to, one or more of starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose and / or aluminum silicate.

[0045] Furthermore, the wetting agent and adhesive include, but are not limited to, one or more of water, glycerol, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia gum slurry, gelatin slurry, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate and / or polyvinyl pyrrolidone.

[0046] Furthermore, the disintegrant includes, but is not limited to, one or more of dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium dodecyl sulfate, methyl cellulose and / or ethyl cellulose.

[0047] Furthermore, the disintegration inhibitor includes but is not limited to sucrose, tristearin, cocoa butter and / or hydrogenated oil.

[0048] Furthermore, the absorption enhancer includes but is not limited to one or more of quaternary ammonium salts and / or sodium lauryl sulfate.

[0049] Furthermore, the lubricant includes, but is not limited to, one or more of talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin and / or polyethylene glycol.

[0050] Furthermore, the tablets can be further made into coated tablets, including sugar-coated tablets, film-coated tablets, enteric-coated tablets, double-layer tablets and multi-layer tablets.

[0051] Furthermore, the pills may be prepared using a wide variety of carriers known in the art, including diluents and absorbents, binders and / or disintegrants.

[0052] Furthermore, the diluent and absorbent include but are not limited to one or more of glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinyl pyrrolidone, Gelucire, kaolin and / or talc.

[0053] Furthermore, the adhesive includes, but is not limited to, one or more of gum arabic, gum tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste and / or batter.

[0054] Furthermore, the disintegrant includes, but is not limited to, one or more of agar powder, dry starch, alginate, sodium dodecyl sulfate, methyl cellulose and / or ethyl cellulose.

[0055] Furthermore, the suppository may be made of a wide variety of carriers known in the art, including but not limited to one or more of polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin and / or semi-synthetic glycerides.

[0056] Furthermore, the injectable preparation includes, but is not limited to, one or more of a solution, an emulsion, a lyophilized powder injection and / or a suspension.

[0057] Furthermore, the injectable preparation may use all diluents commonly used in the art, including but not limited to one or more of water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isostearyl alcohol and / or polyoxyethylene sorbitan fatty acid esters.

[0058] Furthermore, in order to prepare an isotonic injection solution, the injection preparation may be added with an appropriate amount of one or more of sodium chloride, glucose, glycerol, conventional cosolvents, buffers and / or pH regulators.

[0059] Furthermore, the various preparations may also add colorants, preservatives, spices, flavoring agents, sweeteners or other materials to the pharmaceutical preparations as needed.

[0060] Furthermore, the pharmaceutical composition can be administered by injection, cavity administration, respiratory tract administration or mucosal administration.

[0061] Furthermore, the injection administration includes subcutaneous injection, intravenous injection, intramuscular injection and intracavitary injection, etc.; the cavity administration includes rectal or vaginal administration; and the respiratory tract administration includes nasal administration.

[0062] In a fourth aspect, the present invention provides an EGFR inhibitor, wherein the EGFR inhibitor comprises the compound represented by formula (I) of the first aspect, or a pharmaceutically acceptable salt thereof; the compound has the structural formula shown in (I):

[0063]

[0064] Wherein, R is selected from a halogen atom, an aryl group or a heteroaryl group.

[0065] Furthermore, the aromatic group may be a benzene ring or an aromatic hetero group.

[0066] Furthermore, the aryl group and the aromatic hetero group are substituted by one or more substituents.

[0067] Furthermore, the substituent includes, but is not limited to, an alkyl group, a phenolic hydroxyl group or a halogen.

[0068] Furthermore, pharmaceutically acceptable salts of the compound represented by formula (I) include, but are not limited to, hydrochloride, sulfate, maleate, succinate, adipate, glycolate, malate, fumarate, benzenesulfonate, benzoate, hippurate or oxalate.

[0069] Furthermore, the pharmaceutically acceptable salts of the compound represented by formula (I) also include solvates, hydrates or polymorphs of the above salts.

[0070] In a fifth aspect, the present invention provides the use of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a disease caused by EGFR mutation; the structural formula of the compound is shown in (I):

[0071]

[0072] Wherein, R is selected from a halogen atom, an aryl group or a heteroaryl group.

[0073] Furthermore, the aromatic group may be a benzene ring or an aromatic hetero group.

[0074] Furthermore, the aryl group and the aromatic hetero group are substituted by one or more substituents.

[0075] Furthermore, the substituent includes, but is not limited to, an alkyl group, a phenolic hydroxyl group or a halogen.

[0076] Furthermore, pharmaceutically acceptable salts of the compound represented by formula (I) include, but are not limited to, hydrochloride, sulfate, maleate, succinate, adipate, glycolate, malate, fumarate, benzenesulfonate, benzoate, hippurate or oxalate.

[0077] Furthermore, the pharmaceutically acceptable salts of the compound represented by formula (I) also include solvates, hydrates or polymorphs of the above salts.

[0078] Furthermore, the diseases caused by the EGFR mutation include but are not limited to lung adenocarcinoma, lung squamous cell carcinoma, small cell lung cancer and / or non-small cell lung cancer.

[0079] Preferably, the disease caused by the EGFR mutation is non-small cell lung cancer.

[0080] In a sixth aspect, the present invention provides the use of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating cancer; the structural formula of the compound is shown in (I):

[0081]

[0082] Wherein, R is selected from a halogen atom, an aryl group or a heteroaryl group.

[0083] Furthermore, the aromatic group may be a benzene ring or an aromatic hetero group.

[0084] Furthermore, the aryl group and the aromatic hetero group are substituted by one or more substituents.

[0085] Furthermore, the substituent includes, but is not limited to, an alkyl group, a phenolic hydroxyl group or a halogen.

[0086] Furthermore, pharmaceutically acceptable salts of the compound represented by formula (I) include, but are not limited to, hydrochloride, sulfate, maleate, succinate, adipate, glycolate, malate, fumarate, benzenesulfonate, benzoate, hippurate or oxalate.

[0087] Furthermore, the pharmaceutically acceptable salts of the compound represented by formula (I) also include solvates, hydrates or polymorphs of the above salts.

[0088] Furthermore, the cancer includes but is not limited to lung adenocarcinoma, lung squamous cell carcinoma, small cell lung cancer and / or non-small cell lung cancer.

[0089] Preferably, the cancer is non-small cell lung cancer.

[0090] Beneficial Effects

[0091] The present invention starts from the structure of 6,7-dimethoxyquinazoline, and utilizes its 6,7-methoxy groups as hydrogen bond receptors in the common drug design structure to increase affinity with the target. An aromatic group is introduced at the 2-position to adjust the conformation of the compound at the binding site and enhance its binding ability to EGFR containing T790M and C797S mutations. In addition, a piperazine methyl structure is introduced at the 4-position, which, as a pharmaceutical intermediate, can effectively adjust the lipid-water partition coefficient and acid-base equilibrium constant of the drug, thereby improving the biological activity of the drug and its compatibility with the human body, thereby obtaining a 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound.

[0092] In terms of the synthesis method, the present invention increases the flexibility of the compound by inserting a linker with a length of two carbon atoms between the piperazine group and quinazoline-4-amine, thereby better adapting to the binding of important mutant amino acid residues. This design targets the drug resistance problem generated by EGFR-TK inhibitors during treatment and significantly improves the anti-cancer activity of the drug in drug-resistant cells.

[0093] The present invention makes full use of the potential sites of the chemical structure and introduces a piperazine methyl group into the quinazoline skeleton to significantly enhance the inhibitor's activity against EGFR triple mutations, thus filling this gap in the prior art. This innovative design not only expands the activity spectrum of EGFR inhibitors, but also provides new ideas and solutions for overcoming drug resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0094] Figure 1 For intermediate Ⅰ 1 H NMR spectrum.

[0095] Figure 2 For compound 1 1 H NMR spectrum.

[0096] Figure 3 For compound 1 13 C NMR spectrum.

[0097] Figure 4 is the HRMS spectrum of compound 1.

[0098] Figure 5 For compound 2 1 H NMR spectrum.

[0099] Figure 6 For compound 2 13 C NMR spectrum.

[0100] Figure 7 is the HRMS spectrum of compound 2.

[0101] Figure 8 For compound 3 1 H NMR spectrum.

[0102] Fig. 9 For compound 3 13 C NMR spectrum.

[0103] Fig.10 is the HRMS spectrum of compound 3.

[0104] Fig.11 For compound 4 1 H NMR spectrum.

[0105] Fig.12 For compound 4 13 C NMR spectrum.

[0106] Fig.13 is the HRMS spectrum of compound 4. DETAILED DESCRIPTION

[0107] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0108] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0109] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0110] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0111] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0112] All starting materials and solvents are commercially available. All reactions are monitored by thin layer chromatography (TLC) and observed by color development under ultraviolet light (254 or 365nm) or iodine fumigation. All reaction systems are purified. NMR is measured using a Bruker AVANCE III nuclear magnetic spectrometer, and the solvents are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS). All chemical shifts are reported in parts per million (ppm), and the following abbreviations are used to describe the peak splitting mode when necessary: ​​s (singlet), d (doublet), t (triplet), m (multiplet), dd (double doublet). The coupling constant (J) is expressed in Hertz (Hz). MS is measured using LC (Agilent1260Infinity) / MS (G6125B) mass spectrometer (manufacturer: Agilent). High-resolution mass spectrometry (HRMS) was performed using a liquid chromatography-tandem mass spectrometer (XevoG2 Qtof).

[0113] Table 1 Structures of exemplary compounds of the present invention

[0114]

[0115]

[0116] Example 1 Synthesis of 2-chloro-6,7-dimethoxy-N-(2-(4-methylpiperazin-1-yl)ethyl)quinazoline-4-amine (Intermediate I)

[0117] Take a flask, add 1.00g (3.86mmol) of 2,4-dichloro-6,7-dimethoxyquinazoline, dissolve in 5mL of dichloromethane, add 1mL of N,N-diisopropylethylamine, then drop 0.91mL (6.08mmol) of 2-(4-methylpiperazine-1-yl)ethylamine, and stir at 25°C for 48h. After the reaction is completed, dichloromethane and N,N-diisopropylethylamine are removed by vacuum distillation, and silica gel column chromatography is performed to obtain 0.99g of off-white solid with a yield of 70.11%.

[0118] Intermediate I 1 HNMR spectrum see Figure 1 .

[0119] Example 2 Synthesis of 6,7-dimethoxy-N-(2-(4-methylpiperazine-1-yl)ethyl)-2-phenylquinazoline-4-amine

[0120] Weigh 0.2g (0.546mmol) of the intermediate 1 obtained in Example 1, and weigh 86.65mg (0.711mmol) of phenylboronic acid and an appropriate amount of tetrakistriphenylphosphine palladium, add them into a 100mL three-necked flask, replace the air in the flask with nitrogen, add 1.5mL of toluene, 2.5mL of n-butanol, 2.5mL of water and 1mL of triethylamine, control the temperature at 115℃ and stir for 8h. Monitor the reaction by thin layer chromatography and it is basically completed. Stop the reaction, concentrate the insoluble matter in the diatomaceous earth filtration system, separate and purify by column chromatography, the yield is 50.8%; high resolution mass spectrometry: Chemical formula: C 23 H 29 N5O2:408.2394, measured:408.2393.

[0121] Compound 1 1 HNMR spectrum see Figure 2 , 13 C NMR spectrum see Figure 3 , HRMS spectrum see Figure 4 .

[0122] Example 3 Synthesis of 2-(6,7-dimethoxy-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)quinazolin-2-yl)phenol

[0123] The preparation method of 2 is similar to 1, and yellow-green solid 2 is obtained, yield: 80%; high-resolution mass spectrometry: Chemical formula: C 23 H 29 N5O3:424.2343, measured:424.2343.

[0124] Compound 2 1 HNMR spectrum see Figure 5 , 13 C NMR spectrum see Figure 6 , HRMS spectrum see Figure 7 .

[0125] Example 4 Synthesis of 4-chloro-2-(6,7-dimethoxy-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)quinazolin-2-yl)phenol

[0126] The preparation method of 3 is similar to 1, and yellow-green solid 3 is obtained, yield: 52%; high-resolution mass spectrometry: Chemical formula: C 23 H 28 ClN5O3:458.1953, measured:458.1955.

[0127] Compound 4 1 HNMR spectrum see Figure 8 , 13 C NMR spectrum see Fig. 9 , HRMS spectrum see Fig.10 .

[0128] Example 5 Synthesis of 4-fluoro-2-(4-((2-(4-methylpiperazin-1-yl)ethyl)amino)quinazolin-2-yl)phenol

[0129] The preparation method of 4 is similar to 1, and dark yellow solid 4 is obtained, yield: 46.6%; high resolution mass spectrometry: Chemical formula: C 23 H 28 FN5O3:442.2249, measured:442.2288.

[0130] Compound 4 1 HNMR spectrum see Fig.11 , 13 C NMR spectrum see Fig.12 , HRMS spectrum see Fig.11 .

[0131] Example 6 Study on EGFR kinase inhibition concentration of compounds

[0132] All EGFR kinase inhibition experiments were performed using the ADP-Glo ​​kinase kit from Promega. The final compound concentrations were 100 μM, 25 μM, 6.25 μM, 1.5625 μM, 390 nM, 97.7 nM, 24.4 nM, and 0, respectively. Osimertinib was set as a positive control at each concentration. Each compound was mixed with the following components: 0.2 mg / mL substrate (Poly G:T (4:1), from Signal Chem), 5 mmol / L ATP (from Invitrogen), 5 ng EGFR enzyme, and kinase reaction buffer (40 mmol / LTris-HCl, 10 mmol / LMgCl2, and 0.1 mg / mL BSA (bovine serum albumin)) and reacted in a 384-well plate. After reacting at room temperature for 30 min, ADP-Glo ​​reagent (Promega) was added and incubated at room temperature for 40 min. Finally, kinase detection reagent was added and reacted at room temperature for 30 min. Luminescent signals were detected using a microplate ELISA reader (Bio-Tek).

[0133] Data were analyzed using GraphPad Prism. 50 The concentration that causes 50% inhibition of cell growth, ie, the half-inhibitory concentration of antiproliferation, was calculated by a nonlinear regression analysis protocol, ie, "log(inhibitor) vs normalizedresponse—Variable slop".

[0134] The half inhibitory concentration (IC) of osimertinib and the compounds of the present invention is 50 , μM) see Table 2.

[0135] Table 2

[0136]

[0137] In the biological activity experiment of the inhibition of EGFRDel19 / T790M / C797S kinase, the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline derivative prepared by the present invention overcomes the osimertinib resistance and shows good inhibition of the osimertinib-resistant mutant EGFRDel19 / T790M / C797S kinase compared with osimertinib.

[0138] Study on the half-inhibitory concentration of the compound in Example 7

[0139] Cells: PC-9 cell line carrying triple EGFR mutation (EGFR Del19 / T790M / C797S) was cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS). The cells were cultured at 37° C. in a humidified environment containing 5% CO 2 .

[0140] Prepare a cell suspension and count active cells in the logarithmic growth phase. Inoculate the cell suspension (12,000 per well) and incubate at 37°C, 5% CO2 for 12-24 hours. Then remove the residual culture medium and add the medium containing the drug. After 48 hours, remove the old culture medium and add the medium containing 10% CCK-8. Mix well. Monitor color development at 0.5, 1.0 and 2.0 hours. Measure the absorbance at 450nm using a microplate reader. Repeat the experiment three times and calculate the average to obtain the final result. Blank group setup: Add the same volume of CCK-8 as the experimental wells to the 96-well plate, incubate for the same time as the experimental wells, and then measure the absorbance at 450nm.

[0141] Data were analyzed using GraphPad Prism 10.0. 50 The concentration that causes 50% inhibition of cell growth, ie, the half-inhibitory concentration of antiproliferation, was calculated by a nonlinear regression analysis protocol, ie, "log(inhibitor) vs normalizedresponse—Variable slop".

[0142] The half inhibitory concentration (IC 50 , μM) see Table 3.

[0143] Table 3

[0144]

[0145] As can be seen from Table 3, the quinazoline compound of 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline of the present invention has a good inhibitory effect on the proliferation of osimertinib-resistant cell line PC-9 (EGFR Del19 / T790M / C797S), and can be used as an anti-tumor candidate drug after third-generation EGFR-TKI resistance.

Claims

1. A 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound or a pharmaceutically acceptable salt thereof; the compound structural formula is shown in (I): in, R is selected from halogen atoms, aryl groups or heteroaryl groups; the aryl group may be a benzene ring or an aromatic hetero group; the aryl group and the aromatic hetero group are substituted by one or more substituents selected from alkyl groups, phenolic hydroxyl groups or halogens.

2. The compound according to claim 1, wherein the structural formula of the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound can be:

3. A method for preparing the compound according to any one of claims 1 or 2, comprising the steps of: S1. Using 2,4-dichloro-6,7-dimethoxyquinazoline as a raw material, 2-(4-methylpiperazine-1-yl)ethylamine was added to carry out a nucleophilic substitution reaction on the chlorine at the 4 position; S2. Add substituted aryl boronic acid to the reaction system of S1, and use a metal catalyst to carry out a Suzuki coupling reaction to obtain the 6,7-dimethoxy-2-phenyl-4-methylpiperazineethylaminoquinazoline compound.

4. The preparation method according to claim 3, wherein in step S1, the addition ratio of 2,4-dichloro-6,7-dimethoxyquinazoline and 2-(4-methylpiperazine-1-yl)ethylamine is 1:1.2; the nucleophilic substitution reaction temperature is 15-40° C.; the nucleophilic substitution reaction time is 24-48 h; In step S2, the molar ratio of the substituted aryl boronic acid to the metal catalyst is 1:0.01; the Suzuki coupling reaction temperature is 100-130° C.; and the Suzuki coupling reaction time is 5-10 h.

5. The preparation method as claimed in claim 3, in step S2, the substituted aryl boronic acid includes but is not limited to benzo[d][1,3]dioxane-4-yl boronic acid, (5-fluoro-2-hydroxyphenyl)boric acid, 3-furyl boronic acid, (5-chloro-2-hydroxyphenyl)boric acid, 2-hydroxyphenylboric acid, 3-hydroxynaphthalene-2-yl boronic acid, phenylboric acid, 3,5-dichlorophenylboric acid, 2,5-dimethoxyphenylboric acid or 5-cyano-2-methoxyphenylboric acid); the metal catalyst includes but is not limited to tetrakistriphenylphosphine palladium, palladium acetate, dichlorobis(triphenylphosphine)palladium, di(tri-tert-butylphosphine)palladium or palladium on carbon (Pd / C).

6. A pharmaceutical composition comprising a compound as shown in formula (I) of claim 1 or a pharmaceutically acceptable salt thereof and an optional pharmaceutically acceptable carrier; the compound structural formula is shown in (I): in, R is selected from halogen atoms, aryl groups or heteroaryl groups; the aryl group may be a benzene ring or an aromatic hetero group; the aryl group and the aromatic hetero group are substituted by one or more substituents selected from alkyl groups, phenolic hydroxyl groups or halogens.

7. An EGFR inhibitor, comprising a compound as shown in formula (I) of claim 1, or a pharmaceutically acceptable salt thereof; the compound structural formula is shown in (I): in, R is selected from halogen atoms, aryl groups or heteroaryl groups; the aryl group may be a benzene ring or an aromatic hetero group; the aryl group and the aromatic hetero group are substituted by one or more substituents selected from alkyl groups, phenolic hydroxyl groups or halogens.

8. Use of a compound as shown in formula (I) of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a disease caused by EGFR mutation; the disease caused by EGFR mutation includes but is not limited to lung adenocarcinoma, lung squamous cell carcinoma, small cell lung cancer and / or non-small cell lung cancer; the compound structural formula is shown in (I): in, R is selected from halogen atoms, aryl groups or heteroaryl groups; the aryl group may be a benzene ring or an aromatic hetero group; the aryl group and the aromatic hetero group are substituted by one or more substituents selected from alkyl groups, phenolic hydroxyl groups or halogens.

9. Use of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating cancer; the structural formula of the compound is as shown in (I): in, R is selected from halogen atoms, aryl groups or heteroaryl groups; the aryl group may be a benzene ring or an aromatic hetero group; the aryl group and the aromatic hetero group are substituted by one or more substituents selected from alkyl groups, phenolic hydroxyl groups or halogens.

10. The use according to claim 9, wherein the cancer includes but is not limited to lung adenocarcinoma, lung squamous cell carcinoma, small cell lung cancer and / or non-small cell lung cancer, preferably non-small cell lung cancer.

Images